CRISPR-Cas9 Ribonucleoprotein-Mediated Genomic Editing in Primary Innate Immune Cells

STAR Protoc. 2020 Sep 15;1(3):100113. doi: 10.1016/j.xpro.2020.100113. eCollection 2020 Dec 18.

Abstract

CRISPR-Cas9 genome engineering can be used to functionally investigate the complex mechanisms of immune system regulation. Decades of work have aimed to genetically reprogram innate immunity, but current approaches are inefficient or nonspecific, limiting their use. Here, we detail an optimized strategy for non-viral CRISPR-Cas9 ribonucleoprotein (cRNP) genomic editing of primary innate lymphocytes (ILCs) and myeloid lineage cells, resulting in high-efficiency editing of target gene expression from a single electroporation. For complete details on the use and execution of this protocol, please refer to Riggan et al. (2020).

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics
  • Electroporation / methods*
  • Gene Editing / methods*
  • Genomics
  • Humans
  • Immunity, Innate / genetics*
  • Immunity, Innate / physiology
  • Lymphocytes / metabolism
  • Myeloid Cells / metabolism
  • Ribonucleoproteins / genetics
  • Ribonucleoproteins / metabolism

Substances

  • Ribonucleoproteins